Method to produce a highly concentrated immunoglobulin preparation for subcutaneous use

Abstract

The present invention relates to a new and improved method for preparing a highly concentrated immunoglobulin composition from pooled plasma for subcutaneous injection. A composition comprising 20% or more immunoglobulin suitable for subcutaneous use is also described.

What is claimed is: 
     
       1. A method for preparing a concentrated immunoglobulin G (IgG) composition, comprising the steps:
 (A) concentrating a first solution comprising IgG to a protein concentration of from 2% to 10% (w/v) by ultrafiltration using a first ultra-/diafiltration system comprising a first ultrafiltration membrane, thereby forming a first IgG concentrate; 
 (B) diafiltering the first IgG concentrate against a diafiltration buffer using the first ultra-diafiltration system comprising the first ultrafiltration membrane, thereby forming a first IgG diafiltrate; 
 (C) concentrating the first IgG diafiltrate to a protein concentration of greater than 20% (w/v) by ultrafiltration using the first ultra-/diafiltration system comprising the first ultrafiltration membrane, thereby forming a second IgG concentrate; 
 (D) collecting the second IgG concentrate from the first ultra-/diafiltration system; 
 (E) washing the first ultrafiltration membrane by re-circulating a post-wash buffer through the first ultra-/diafiltration system wherein the first ultra-/diafiltration system is washed with a volume of post-wash buffer equal to at least two times the dead volume of the first ultra-/diafiltration system, thereby forming a first IgG post-wash solution; 
 (F) transfering the first IgG post-wash solution from the first ultra-/diafiltration system into a second ultra-/diafiltration system comprising a second ultrafiltration membrane, wherein the surface area of the second ultrafiltration membrane is lower than the surface area of the first ultrafiltration membrane; 
 (G) concentrating the first IgG post-wash solution to a protein concentration of greater than 20% (w/v) by ultrafiltration using the second ultra-/diafiltration system comprising a second ultra-/diafiltration membrane, thereby forming a third IgG concentrate; and 
 (H) combining the third IgG concentrate from the second ultra-/diafiltration system with the second IgG concentrate, thereby forming a concentrated IgG composition. 
 
     
     
       2. The method of  claim 1 , wherein the first solution comprising IgG is concentrated in (A) to a protein concentration of 5±1% (w/v). 
     
     
       3. The method of  claim 1 , wherein the first ultrafiltation membrane is an open-screen membrane. 
     
     
       4. The method of  claim 1 , wherein the first or second ultrafiltration membrane have a nominal molecular weight cut off (NMWCO) of 100 kDa or less. 
     
     
       5. The method of  claim 1 , wherein the first and second ultrafiltration membrane has a nominal molecular weight cut off (NMWCO) of 100 kDa or less. 
     
     
       6. The method of  claim 1 , wherein the first or second ultrafiltration membrane has a nominal molecular weight cut off (NMWCO) of 90 kDa or less. 
     
     
       7. The method of  claim 1 , wherein the first and second ultrafiltration membrane have a nominal molecular weight cut off (NMWCO) of 90 kDa or less. 
     
     
       8. The method of  claim 1 , wherein the first and second ultrafiltration membrane have a same nominal molecular weight cut off (NMWCO). 
     
     
       9. The method of  claim 1 , wherein the diafiltration buffer comprises from 0.2 M to 0.3 M glycine and a pH of 4.2±0.1. 
     
     
       10. The method of  claim 1 , wherein the second IgG concentrate has a protein concentration of at least 22% (w/v). 
     
     
       11. The method of  claim 1 , wherein the surface area of the second ultrafiltration membrane is no more than a tenth of the surface area of the first ultrafiltration membrane. 
     
     
       12. The method of  claim 1 , wherein the protein concentration of the concentrated IgG composition formed in (H) is greater than 20% (w/v), and wherein the method further comprises adjusting the concentration of the concentrated IgG composition formed in (H) to about 20% (w/v). 
     
     
       13. The method of  claim 1 , wherein the pH of the concentrated IgG composition formed in (H) is from 4.0 to 6.0. 
     
     
       14. The method of  claim 1 , further comprising the steps of:
 (I) washing the second ultrafiltration membrane by re-circulating a post-wash buffer through the second ultra-/diafiltration system, thereby forming a second IgG post-wash solution; and 
 (J) adjusting the protein concentration of the concentrated IgG composition formed in (H) by admixing at least a portion of the second IgG post-wash solution into the concentrated IgG composition formed in (H). 
 
     
     
       15. The method of  claim 1 , wherein the IgG is human IgG. 
     
     
       16. The method of  claim 1 , comprising the steps:
 (A) concentrating a first solution comprising IgG to a protein concentration of 5±1% (w/v) by ultrafiltration using a first ultra-/diafiltration system comprising a first ultrafiltration membrane, thereby forming a first IgG concentrate, the first ultrafiltration membrane having a NMWCO of 60 kDa or less; 
 (B) diafiltering the first IgG concentrate against a diafiltration buffer using the first ultra-/diafiltration system comprising the first ultrafiltration membrane, thereby forming a first IgG diafiltrate, the diafiltration buffer comprising glycine and a pH of 4.2±0.1; 
 (C) concentrating the first IgG diafiltrate to a protein concentration of greater than 20% (w/v) by ultrafiltration using the first ultra-/diafiltration system comprising the first ultrafiltration membrane, thereby forming a second IgG concentrate; 
 (D) collecting the second IgG concentrate from the first ultra-/diafiltration system; 
 (E) washing the first ultrafiltration membrane by re-circulating a post-wash buffer through the first ultra-/diafiltration system, thereby forming a first IgG post-wash solution, the post-wash buffer having a volume equal to at least two times the dead volume of the first ultra-/diafiltration system; 
 (F) collecting the first IgG post-wash solution from the first ultra-/diafiltration system into a second ultra-/diafiltration system comprising a second ultrafiltration membrane, the second ultrafiltration membrane having a NMWCO of 60 kDa or less; 
 (G) concentrating the first IgG post-wash solution to a protein concentration of greater than 20% (w/v) by ultrafiltration using the second ultra-/diafiltration system comprising a second ultra-/diafiltration membrane, wherein the surface area of the second ultrafiltration membrane is no more than a tenth of the surface area of the first ultrafiltration membrane, thereby forming a third IgG concentrate; 
 (H) combining the third IgG concentrate from the second ultra-/diafiltration system with the second IgG concentrate, thereby forming a concentrated IgG composition; 
 (I) washing the second ultrafiltration membrane by re-circulating a post-wash buffer through the second ultra-/diafiltration system, thereby forming a second IgG post-wash solution; and 
 (J) adjusting the protein concentration of the concentrated IgG composition formed in (H) by admixing at least a portion of the second IgG post-wash solution into the concentrated IgG composition formed in (H).